Created of Warm Blood and Nerves: Restoring an Enteric Nervous System in Organoids  Christopher R. Schlieve, Tracy C. Grikscheit  Cell Stem Cell  Volume 20, Issue 1, Pages 5-7 (January 2017) DOI: 10.1016/j.stem.2016.12.012 Copyright © 2017 Elsevier Inc. Terms and Conditions

Figure 1 Co-culture of Human Intestinal Organoids with Enteric Neural Crest Cells Restores Key Elements of the Enteric Nervous System In Vitro and In Vivo The diagram demonstrates the approach for establishing a human model for studying ENS-intestinal biology derived from hESCs or hiPSCs. HIOs in vitro without ENCC supplementation develop a rudimentary epithelium and underlying mesenchyme. ENCCs combined with HIOs for 4 weeks in vitro migrate within the muscular layer, differentiate into neurons and glia, and demonstrate neuronal activity through calcium imaging. Excitatory and interneurons were identified after in vitro growth, but an absence of inhibitory nNOS-positive neurons was observed. In vivo, enteric neurons and glia coalesced to form immature myenteric ganglia and submucosal ganglia with smaller nerve bundles than adult intestine. HIOs + ENS failed to restore the neuroepithelial network between sensory neurons and enteroendocrine cells and would therefore lack the necessary connectivity required for intraluminal sensing and signaling to the mesenchyme and ENS. Taken together, this method restores some aspects of an immature ENS, representative of fetal intestinal development. Cell Stem Cell 2017 20, 5-7DOI: (10.1016/j.stem.2016.12.012) Copyright © 2017 Elsevier Inc. Terms and Conditions